6-His-MBP-TEV-FnCpf1 vector (V010156)

Basic Vector Information

Gene/Insert name FnCpf1 (humanized) Alt name type V CRISPR-associated protein Cpf1 (humanized) Alt name Cpf1 (humanized) Alt name Francisella tularensis subsp. novicida U112 Cpf1 (humanized) Species Francisella tularensis subsp. novicida U112 Insert Size (bp) 3910 Promoter T7 Tags / Fusion Proteins 6xHis (N terminal on backbone) MBP (N terminal on insert) TEV site (N terminal on insert) Nucleoplasmin NLS (C terminal on insert) 3xHA (C terminal on insert)

      • Vector Name:
      • 6-His-MBP-TEV-FnCpf1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 10685 bp
      • Type:
      • CRISPR Plasmids
      • Source/Author:
      • Feng Zhang
      • Copy Number:
      • High copy number
      • Promoter:
      • T7
      • Cloning Method:
      • Enzyme Cut
      • 5' Primer:
      • T7 promoter
      • 3' Primer:
      • T7 terminator
      • Fusion Tag:
      • MBP
      • Expression Method:
      • IPTG induced

6-His-MBP-TEV-FnCpf1 vector Vector Map

6-His-MBP-TEV-FnCpf110685 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500oribomropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBS6xHisMBPthrombin siteTEV sitecas12anucleoplasmin NLS3xHAFLAGAviTag(TM)6xHisT7 terminatorM13 oriAmpR promoterAmpR

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

6-His-MBP-TEV-FnCpf1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       6-His-MBP-TEV-Fn       10685 bp DNA     circular SYN 04-JUL-2018
DEFINITION  Bacterial expression plasmid for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    6-His-MBP-TEV-FnCpf1.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10685)
  AUTHORS   Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, 
            Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin 
            EV, Zhang F
  TITLE     Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas 
            System.
  JOURNAL   Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 
            10.1016/j.cell.2015.09.038.
  PUBMED    26422227
REFERENCE   2  (bases 1 to 10685)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10685)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2015 
            Sep 23. pii: S0092-8674(15)01200-3. doi: 
            10.1016/j.cell.2015.09.038."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      532..1120
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1306..1445)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(1550..1738)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(2513..2534)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2550..3629)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3630..3707)
                     /label=lacI promoter
     promoter        4016..4034
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4035..4059
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             4074..4096
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4110..4127
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             4128..5225
                     /label=MBP
                     /note="maltose binding protein from E. coli"
     CDS             5244..5261
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             5268..5288
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             5289..9188
                     /gene="cas12a"
                     /label=cas12a
                     /note="CRISPR-associated endonuclease Cas12a from
                     Francisella tularensis subsp. novicida (strain U112). 
                     Accession#: A0Q7Q2"
     CDS             9189..9236
                     /label=nucleoplasmin NLS
                     /note="bipartite nuclear localization signal from
                     nucleoplasmin"
     CDS             9243..9323
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     CDS             9348..9371
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             9372..9416
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     CDS             9432..9449
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      9516..9563
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      9600..9980
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        10081..10185
                     /label=AmpR promoter
     CDS             join(10186..10685,1..358)
                     /label=AmpR
                     /note="beta-lactamase"

This page is informational only.